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Background and Aims: Leakage rates from colonic anastomoses continue to be significant. The effect of luminal butyrate on the healing of colonic anastomoses in the ascending and descending colon was investigated in rats. Methods: Colonic resection and anastomosis was performed in the proximal and distal colon of adult Wistar rats. Daily enemas of butyrate (40 mM) or saline were administered to case and control rats respectively. Animals were sacrificed on the 6th day and colonic segments of 4 cm including the anastomoses were excised from both the ascending and descending colons. Bursting pressure for the anastomotic line was measured for each colonic segment. Results: The mean bursting pressure was significantly higher (131.96±20.11 mm Hg, mean+SD) in the group receiving butyrate enemas as compared to the control group receiving saline enemas (91.29±15.91) (P<0.001). The effect of butyrate on bursting wall tension was higher in the proximal colon than in the distal colon. Conclusion: Butyrate facilitates colonic anastomotic healing and results in significantly stronger anastomoses judged by the bursting pressure. There may be a role for local butyrate instillation in healing of colonic anastomoses.